WO2023171463A1 - Dispositif de traitement d'informations et système de traitement d'informations - Google Patents

Dispositif de traitement d'informations et système de traitement d'informations Download PDF

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WO2023171463A1
WO2023171463A1 PCT/JP2023/007273 JP2023007273W WO2023171463A1 WO 2023171463 A1 WO2023171463 A1 WO 2023171463A1 JP 2023007273 W JP2023007273 W JP 2023007273W WO 2023171463 A1 WO2023171463 A1 WO 2023171463A1
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Prior art keywords
information
single staining
information processing
fluorescent dye
staining spectrum
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PCT/JP2023/007273
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English (en)
Japanese (ja)
Inventor
孝治 二村
旺秀 樋口
史高 大塚
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ソニーグループ株式会社
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Publication of WO2023171463A1 publication Critical patent/WO2023171463A1/fr

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence

Definitions

  • the present disclosure relates to an information processing device and an information processing system. More specifically, the present disclosure relates to an information processing device and an information processing system that perform processing on spectral data of fluorescent dyes.
  • a particle population such as cells, microorganisms, and liposomes is labeled with a fluorescent dye, and each particle in the particle group is irradiated with laser light to measure the intensity and/or pattern of fluorescence generated from the excited fluorescent dye. This is used to measure the properties of particles.
  • a typical example of a particle analyzer that performs this measurement is a flow cytometer.
  • a flow cytometer irradiates particles flowing in a line in a flow channel with laser light (excitation light) of a specific wavelength and detects the fluorescence and/or scattered light emitted from each particle. , is a device that analyzes multiple particles one by one.
  • a flow cytometer converts the light detected by a photodetector into an electrical signal, digitizes it, and performs statistical analysis to determine the characteristics of individual particles, such as type, size, and structure. can.
  • a combination of one or more fluorescent dye-labeled antibodies is used to perform flow cytometric analysis.
  • This combination design is also called a panel design.
  • panel design several techniques have been proposed so far.
  • Patent Document 1 discloses a reagent selection support device that supports selection of reagents used for cell measurement.
  • the reagent selection support device includes: an acquisition unit that acquires order information including a plurality of measurement items; a first fluorescent reagent used to measure a first target molecule corresponding to the plurality of measurement items; The combination of the second fluorescent reagent used to measure the second target molecule corresponding to the second measurement item different from the measurement item of a processing unit that determines based on information that reflects the characteristics of the first fluorescent dye, information that reflects the characteristics of the second target element, and the characteristics of the second fluorescent dye contained in the second fluorescent dye; and an output unit that outputs a combination of the first fluorescent reagent and the second fluorescent reagent.
  • Patent Document 2 describes fluorescence generated from fluorescent dyes excited by irradiating light onto microparticles that are multiple-labeled with multiple fluorescent dyes with overlapping fluorescence wavelength bands. is received by photodetectors with different receiving wavelength bands, which are arranged in a number greater than the number of fluorescent dyes, and the measurement spectrum obtained by collecting the detected values from each photodetector is obtained by labeling each fluorescent dye individually.
  • a fluorescence intensity correction method is disclosed that includes a procedure of approximation by a linear sum of single staining spectra obtained with microparticles.
  • JP2019-203842A Japanese Patent Application Publication No. 2011-232259
  • Fluorescent reagents may change over time depending on storage conditions (e.g. light, oxidation, temperature, etc.). Therefore, master data stored as spectrum information of each fluorescent reagent or spectrum information acquired in the past may not reflect the current state of the fluorescent reagent. Utilizing spectral information that does not reflect the current state of the fluorescent reagents may prevent accurate panel design or performing an unmixing process.
  • tandem reagents in particular may have different spectral information even if they are from the same lot but in different bottles. Therefore, especially regarding tandem reagents, master data or previously acquired spectral information may not be suitable for use as current spectral information. Additionally, if the fluorescent reagents deteriorate, the fluorescent reagents in stock may become unusable.
  • an object of the present disclosure is to provide a technique for appropriately understanding the state of a fluorescent reagent.
  • the present disclosure provides an information processing device including a processing section configured to detect a change in single staining spectrum information of a fluorescent dye.
  • the processing unit may compare the first single staining spectrum information and the second single staining spectrum information of the fluorescent dye, and determine whether the single staining spectrum information has changed based on the comparison.
  • the first single staining spectrum information and the second single staining spectrum information are single staining spectrum information regarding the same fluorescent dye, but may be acquired at different times.
  • the single staining spectrum information may be fluorescence intensity spectrum data of fluorescence generated by irradiating the fluorescent dye with light.
  • the single staining spectrum information may be spectrum data represented by fluorescence intensity at each wavelength in a predetermined wavelength range.
  • the processing unit acquires change amount data representing a change between the first single staining spectrum information and the second single staining spectrum information of the fluorescent dye, and based on the change amount data and the comparison. It can be determined whether the single staining spectrum information has changed.
  • the processing unit may determine that the single stain spectrum information has changed when the change amount data is greater than or equal to a predetermined threshold.
  • the processing unit may generate information regarding the change in response to determining that the single staining spectrum information has changed.
  • the information regarding the change may include an alert indicating that the single stain spectral information has changed.
  • the information regarding the change may include information specifying a fluorescent dye whose single staining spectrum information has changed.
  • the information regarding the change may include single staining spectrum graph data of a fluorescent dye whose single staining spectrum information has changed.
  • the processing unit may output a screen including a display prompting the user to select whether to update single staining spectrum information.
  • the processing unit may be configured to perform panel design processing using the updated single staining spectrum information.
  • the processing unit may be configured to perform an unmixing process using the updated single staining spectrum information.
  • the present disclosure also provides an information processing system including a processing unit configured to detect a change in single staining spectral information of a fluorescent dye.
  • the information processing system includes: an information processing device having the processing section;
  • a biological sample analyzer may include.
  • the biological sample analyzer can perform an analysis process using the fluorescent dye.
  • the biological sample analysis device may be configured to transmit a single staining spectrum of the fluorescent dye obtained by the analysis process to the information processing device.
  • FIG. 1 is a diagram illustrating a configuration example of an information processing device according to the present disclosure.
  • FIG. 2 is a flow diagram of an example of a process executed by an information processing device according to the present disclosure. It is a figure showing a typical example of fluorescence spectrum plot data.
  • FIG. 6 is a diagram illustrating an example of a screen that includes information regarding changes.
  • FIG. 7 is a diagram illustrating another example of a screen that includes information regarding changes.
  • FIG. 7 is a diagram illustrating yet another example of a screen that includes information regarding changes. It is a figure which shows the example of the screen which prompts the update of single staining spectrum information.
  • FIG. 7 is a diagram showing another example of a screen that prompts updating of single staining spectrum information.
  • FIG. 1 is a diagram showing an example of the configuration of a biological sample analyzer. 1 is a diagram illustrating a configuration example of an information processing system according to the present disclosure.
  • First embodiment information processing device
  • Overview of the present disclosure (2) Configuration example of an information processing device (3)
  • Example of a process executed by a processing unit (4)
  • Configuration example of a biological sample analyzer (5)
  • Second embodiment information processing system
  • Third embodiment information processing method
  • Fourth embodiment program
  • the present disclosure provides an information processing device that includes a processing section configured to detect changes in single staining spectrum information of a fluorescent dye. By detecting changes in single staining spectrum information, it is possible to prompt the user to select appropriate single staining spectrum information, for example.
  • the fluorescent dye may be a fluorescent dye that constitutes a fluorescent dye-labeled antibody.
  • the fluorescent dye-labeled antibody may be a fluorescent dye-labeled antibody used for staining particles (particularly biological particles such as cells or liposomes). That is, the processing section may be configured to detect a change in the single staining spectrum information of the fluorescent dye-labeled antibody.
  • the change may be, for example, a change in the single staining spectrum information of one or more specific fluorescent dyes, and particularly may be a change over time.
  • the fluorescence emitted by fluorescent dyes can change over time. According to the present disclosure, it is possible to identify changes over time in single staining spectrum information, thereby allowing more appropriate panel design processing or fluorescence correction processing (particularly unmixing processing) to be performed.
  • appropriate single staining spectrum information can be used in various treatments. Utilizing appropriate single staining spectral information enables, for example, appropriate panel design construction or appropriate unmixing processing. Further, the processing unit may prompt the user to select an optimal single staining spectrum for each fluorescent reagent in response to the detection of the change. This contributes to more accurate unmixing processing or panel design construction. Furthermore, by detecting changes in single staining spectrum information, it is also possible to evaluate changes in the fluorescent labeling reagent.
  • the figure is a block diagram of the information processing device.
  • the information processing device 100 shown in the figure may include a processing section 101, a storage section 102, an input section 103, an output section 104, and a communication section 105.
  • the information processing device 100 may be configured as a so-called computer, for example, and may be a desktop, laptop, or tablet computer, or may be a server computer.
  • the processing unit 101 is configured to be able to detect changes in the single staining spectrum information of the fluorescently labeled antibody. More specific processing of the detection will be described in detail below.
  • the processing unit 101 may include, for example, a CPU (Central Processing Unit) and a RAM.
  • the CPU and RAM may be interconnected via a bus, for example.
  • An input/output interface may be further connected to the bus.
  • An input section 103, an output section 104, and a communication section 105 may be connected to the bus via the input/output interface.
  • the storage unit 102 stores various data.
  • the storage unit 102 stores, for example, data acquired in the process described below (for example, first and second single staining spectrum information and information regarding changes, etc.) and/or data generated in the process described below (for example, first and second
  • the storage device may be configured to be able to store information such as single staining spectrum information and information regarding changes, and may include, for example, an information recording medium.
  • these data include various data received by the input unit 103 (for example, the above single stain spectrum information, etc.), various data received via the communication unit 105 (for example, the above single stain spectrum information, etc.), and the processing unit 101 Examples include, but are not limited to, various data generated by (for example, the above-mentioned single staining spectrum information and information regarding changes).
  • the storage unit 102 also includes an operating system (for example, WINDOWS (registered trademark), UNIX (registered trademark), LINUX (registered trademark), etc.), an information processing method according to the present disclosure applied to an information processing device or an information processing system. A program to be executed and various other programs may be stored.
  • the single staining spectrum information (first single staining spectrum information and second single staining spectrum information) stored in the storage unit 102 is based on the date and time data when the single staining spectrum information was acquired (especially for acquiring the single staining spectrum information). (data specifying the date and time when the analysis process was performed) or may include the date and time data. This enables time-series management of single staining spectrum information. It is also possible to specify the date and time when the state of the fluorescent dye was analyzed.
  • the input unit 103 may include an interface configured to accept input of various data.
  • the input unit 103 may be configured to be able to receive various data input in processing described below.
  • the input unit 103 may include, for example, a mouse, a keyboard, a touch panel, and the like as devices that accept such operations.
  • the output unit 104 may include an interface configured to output various data.
  • the output unit 104 may be configured to output various data generated in the processing described below. Examples of the data include, but are not limited to, various data generated by the processing unit 101.
  • the output unit 104 may include, for example, a display device as a device that outputs these data.
  • the communication unit 105 may be configured to connect the information processing device 100 to a network by wire or wirelessly.
  • the communication unit 105 allows the information processing apparatus 100 to acquire various data via the network.
  • the acquired data can be stored in the storage unit 102, for example.
  • the configuration of the communication unit 105 may be appropriately selected by those skilled in the art.
  • the information processing device 100 may include, for example, a drive (not shown).
  • the drive can read data (for example, the various data mentioned above) or programs (such as the programs mentioned above) recorded on the recording medium and output it to the RAM.
  • the recording medium is, for example, a microSD memory card, an SD memory card, or a flash memory, but is not limited to these.
  • FIG. 2 An example of processing executed by the information processing device according to the present disclosure will be described below with reference to FIG. 2.
  • the figure is a flow diagram of the processing example.
  • Step S11 the information processing device 100 (particularly the processing unit 101) starts a process of detecting a change in single staining spectrum information.
  • This process may be performed before starting the panel design process, during the panel design process, or after the panel design process, as will be described later.
  • the process may be performed, for example, before the start of the analysis process by a biological sample analyzer such as a flow cytometer, during the analysis process, or after the end of the analysis process.
  • Step S12 the information processing device 100 acquires single staining spectrum information of the fluorescent dye-labeled antibody.
  • a biological sample analyzer such as a flow cytometer acquires single-staining spectrum information by performing analysis processing on a sample (particularly a single-stained sample) stained with a fluorescent dye-labeled antibody.
  • the information processing device 100 acquires the single staining spectrum information from the biological sample analysis device.
  • Single staining spectrum information may be fluorescence intensity spectrum data of fluorescence generated by irradiation of excitation light to particles labeled with a single fluorescent dye (particularly particles labeled with a single fluorescent dye-labeled antibody). .
  • the single staining spectrum information may be data indicating the fluorescence distribution of a fluorescent dye, and more specifically may be spectrum data represented by fluorescence intensity at each wavelength in a predetermined wavelength range.
  • the fluorescence may be, for example, fluorescence detected by a photodetector.
  • the fluorescence intensity spectral data may be, for example, spectral data generated by combining fluorescence intensity data acquired by each of a plurality of photodetectors having different reception wavelength bands.
  • the single staining spectrum information acquired in step S12 may be treated as fluorescence intensity spectrum data indicating the current state of the fluorescent dye-labeled antibody. It may refer to fluorescence intensity spectral data indicating the current state of the labeled antibody.
  • the single staining spectrum information is also referred to as "second single staining spectrum information.”
  • the information processing apparatus 100 can also acquire reference single staining spectrum information of the fluorescent dye-labeled antibody from which the single staining spectrum information is derived.
  • the reference single staining spectrum information is also referred to as "first single staining spectrum information.” Note that the information processing apparatus 100 may previously hold the reference single staining spectrum information prior to step S12. That is, the first single staining spectrum information and the second single staining spectrum information are single staining spectrum information regarding the same fluorescent dye, but may be obtained at different times.
  • the information processing device 100 stores fluorescent dye identification information or fluorescent dye labeled antibody identification information included in the second single staining spectrum information (or fluorescent dye identification information or fluorescent dye labeled antibody identification information associated with the second single staining spectrum information). information), the first single staining spectrum information may be acquired.
  • the fluorescent dye identification information is information for specifying the fluorescent dye from which the second single staining spectrum information is derived, and may be, for example, the name of the fluorescent dye or a number (such as a product number) for specifying the fluorescent dye.
  • the fluorescent dye-labeled antibody identification information is information for specifying the fluorescent dye-labeled antibody from which the second single staining spectrum information is derived, such as the name of the fluorescent dye-labeled antibody or the number that specifies the fluorescent dye-labeled antibody. (For example, product number, etc.)
  • the information processing apparatus 100 can acquire the first single staining spectrum (reference single staining spectrum) of the fluorescent dye or fluorescent dye-labeled antibody specified by each piece of identification information. Thereby, the information processing apparatus 100 can, for example, automatically acquire the first single staining spectrum, and can save the user the trouble of selecting the first single staining spectrum.
  • the first single staining spectrum information is obtained by using the same fluorescent dye (in particular, a fluorescent dye labeled antibody) as the fluorescent dye (especially a fluorescent dye labeled antibody) that stained the sample used to obtain the second single staining spectrum information. It was obtained by the above-mentioned analysis process on a stained sample.
  • the first single staining spectrum information and the second single staining spectrum information are for a sample containing the same particles (for example, cells of the same type) stained with the same fluorescent dye (particularly a fluorescent dye-labeled antibody). This is obtained by performing the above analysis process on a sample containing different particles (for example, different types of cells) stained with the same fluorescent dye (in particular, a fluorescent dye-labeled antibody). It may be obtained.
  • the cells of the same type may preferably be cells obtained by the same treatment procedure, but may also be cells obtained by different treatment procedures.
  • the analysis processing for obtaining the first single staining spectrum information and the analysis processing for obtaining the second single staining spectrum information are preferably the same, but may be different. Differences in processing procedures and analysis processes can be compensated for by appropriately adjusting the acquired data.
  • the first single staining spectrum information and the second single staining spectrum information may be obtained at different times when the analysis process for obtaining each single staining spectrum information is executed.
  • the first single staining spectrum information may be data acquired in the past than the second single staining spectrum information.
  • the first single staining spectrum information is so-called master data regarding a fluorescent dye (in particular, a fluorescent dye-labeled antibody) that stained the sample used to obtain the second single staining spectrum information. It's okay.
  • a fluorescent dye in particular, a fluorescent dye-labeled antibody
  • various types of processing such as panel design processing or unmixing processing
  • step S13 the information processing device 100 (particularly the processing section) determines whether the single staining spectrum information has changed. In the determination, the information processing apparatus 100 may, for example, compare the first single staining spectrum information and the second single staining spectrum information. The information processing apparatus 100 may then determine whether the single staining spectrum information has changed based on the comparison. For example, the information processing device 100 can compare the fluorescence intensity at each wavelength of the single staining spectrum information.
  • the first single staining spectrum information and the second single staining spectrum information are single staining spectrum information regarding the same fluorescent dye, and in particular are single staining spectrum information of particles stained with the same fluorescent dye labeled antibody.
  • the first single staining spectral information may also be referred to as reference spectral information. That is, in step S13, the information processing apparatus 100 compares the first single staining spectrum information and the second single staining spectrum information, which serve as a reference.
  • the information processing apparatus 100 may perform the comparison based on the fluorescence intensity in a predetermined wavelength range of the first single staining spectrum information and the second single staining spectrum information.
  • the predetermined wavelength range may be the entire wavelength range common to these two single stain spectra, or may be a part of the wavelength range common to these two single stain spectra.
  • the partial wavelength range may be, for example, a wavelength range that covers the maximum fluorescence wavelength.
  • the information processing apparatus 100 can acquire change amount data representing a change between the first single staining spectrum information and the second single staining spectrum information. Fluorescence intensity change amount data at each wavelength may be acquired.
  • the change amount data may be, for example, error data or calculation data of the fluorescence intensity at each wavelength of these two single staining spectrum information (for example, difference data or quotient data of the two spectrum information, or calculation results based on either of these) data).
  • the change amount data is greater than or equal to a predetermined threshold
  • the information processing device 100 may determine that the single staining spectrum information has changed.
  • the change amount data is less than or equal to a predetermined threshold, the information processing apparatus 100 may determine that the single staining spectrum information has not changed.
  • the threshold value may be set as appropriate depending on the type of fluorescent dye. Further, the threshold value may be changed as appropriate depending on, for example, the number of fluorescent dyes included in a panel created in panel design processing. In this way, the information processing device 100 (particularly the processing unit) acquires change amount data representing a change between the first single staining spectrum information and the second single staining spectrum information of the fluorescent dye, and , may be configured to determine whether the single stain spectrum information has changed based on the comparison based on the change amount data.
  • the change amount data is, for example, the difference between "the fluorescence intensity at a certain wavelength of the first single staining spectrum information" and "the fluorescence intensity at the certain wavelength of the second single staining spectrum information".
  • the amount of change data may be based on .
  • the change amount data based on the difference may be expressed by, for example, Expression 1 or Expression 2 below, but is not limited to these.
  • the change amount data may be expressed as an absolute value.
  • the threshold value may be, for example, any value between 0.01 and 0.2, particularly any value between 0.02 and 0.15.
  • the threshold values are, for example, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0. .13, 0.14, or 0.15, but are not limited to these.
  • the threshold value may be, for example, any value between 1% and 20%, particularly any value between 2% and 15%.
  • the threshold value is, for example, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, or 15%. , but not limited to.
  • the single staining spectrum information may be expressed as fluorescence spectrum plot data (or graph data) in which the horizontal axis is the detector channel number and the vertical axis is the fluorescence intensity.
  • a schematic example of the graph data is shown in FIG.
  • the vertical axis represents fluorescence intensity in arbitrary units
  • the horizontal axis represents the channel number of the detector.
  • 1st is first single staining spectrum information
  • 2nd is second single staining spectrum information.
  • single stain spectral information may be visually represented by plotting the fluorescence intensity at each channel number.
  • change amount data can be acquired from the fluorescence intensity at each wavelength of these two single staining spectrum information.
  • the information processing device 100 may change the single staining spectrum information. It can be determined that it did. Further, if there is no wavelength within the predetermined wavelength range for which the change amount data is equal to or greater than a predetermined threshold, the information processing device 100 determines that the single staining spectrum information has not changed. It's fine.
  • the information processing device 100 may perform a single staining spectrum It may be determined that the information has changed. Furthermore, when the average value of the change amount data within the predetermined wavelength range is less than or equal to a predetermined threshold, the information processing device 100 may determine that the single staining spectrum information has not changed. .
  • step S13 if the information processing device 100 determines that the single staining spectrum information has changed, the information processing device 100 advances the process to step S14. Further, when the information processing apparatus 100 determines that the single staining spectrum information has not changed, the information processing apparatus 100 advances the process to step S17.
  • step S14 the information processing device 100 (particularly the processing section) generates information regarding changes in single staining spectrum information.
  • the information processing device 100 outputs information regarding the change.
  • the information processing apparatus 100 may display a screen including information regarding the change on the display device.
  • the information regarding the change may include, for example, alert information indicating that the single staining spectrum information has changed.
  • the information processing device 100 can display an alert display on the display device based on the alert information.
  • the information regarding the change may include, for example, information specifying the fluorescent dye or fluorescent dye-labeled antibody whose single staining spectrum information has changed.
  • the information processing device 100 can display the fluorescent dye or fluorescent dye-labeled antibody with changed single staining spectrum information on the display device based on the specified information. Further, the information regarding the change may include, for example, single staining spectrum graph data of a fluorescent dye or a fluorescent dye labeled antibody whose single staining spectrum information has changed. Based on the graph data, the information processing device 100 can display on the display device a single staining spectrum graph of a fluorescent dye or a fluorescent dye labeled antibody in which single staining spectrum information has changed. In this way, the information processing apparatus 100 may be configured to generate information regarding the change in response to determining that the single staining spectrum information has changed.
  • FIG. 4A An example of a screen containing information regarding the change is shown in FIG. 4A.
  • the above-mentioned alert is displayed as "The following dyes may have changed or deteriorated.”
  • the screen may display a dye whose single staining spectrum information has been determined to have changed.
  • two dyes, "FITC” and "PerCP-Cy5.5”, are shown to be possibly changed or degraded.
  • the screen including information regarding the change may have a display prompting the user to select whether to update the single staining spectrum information.
  • the message "If you want to update the spectrum data, please press the update button” is displayed, but the displayed content is not limited to this.
  • an update button 201 for updating single staining spectrum information is displayed.
  • a Close button 202 for ending the process without updating is displayed in the figure.
  • the information processing apparatus 100 can output a screen including a display prompting the user to select whether to update single staining spectrum information (in particular, can display it on a display device).
  • the display may be displayed in a screen containing information about the change, as described above, or it may be displayed in another screen.
  • FIG. 4B Another example of a screen containing information regarding the change is shown in FIG. 4B.
  • the screen may display a combination of molecules to be captured by the antibody and the fluorescent dye that constitute the fluorescent dye-labeled antibody.
  • a combination of the surface marker "CD235a”, which is a molecule to be captured by the antibody constituting the fluorescent dye-labeled antibody, and the dye "FICT” is displayed as a fluorescent dye-labeled antibody with a changed single staining information spectrum.
  • the combination of the surface marker "CD59” and the dye "PerCP-Cy5.5” is also displayed as a fluorescent dye-labeled antibody whose single staining information spectrum has changed or deteriorated.
  • FIG. 4C Another example of a screen containing information regarding the change is shown in FIG. 4C.
  • graph data of a single staining spectrum of a fluorescent dye may be displayed on the screen.
  • the fluorescent dye-labeled antibody "PerCP-Cy5.5” for which a change in the single staining spectrum was detected and its single staining spectrum data graph 203 are displayed.
  • the graph data displays a "reference spectrum” corresponding to the first single staining spectrum information and a "single staining spectrum” corresponding to the second single staining spectrum information.
  • These two pieces of single staining spectrum information may be displayed in a superimposed manner in one graph data, as shown in the figure. This makes it easy to understand changes in spectral information.
  • the graph data is a graph in which the vertical axis is the fluorescence intensity and the horizontal axis is the fluorescence wavelength
  • the settings of the axes are not limited to these and may be appropriately selected by those skilled in the art.
  • the format of the graph is not limited to the spectral waveform graph shown in the figure, but may be a graph of other formats.
  • step S15 the information processing device 100 determines whether an instruction to change the single staining spectrum information has been received. This determination may be made depending on whether the update button 201 in the figure has been pressed. In response to the selection of the update button 201 shown in the figure, the information processing apparatus 100 advances the process to step S16. In response to selection of the Close button 202 shown in the figure, the information processing device 100 advances the process to step S17.
  • Step S16 the information processing device 100 can display a screen on the display device that prompts the user to update the single staining spectrum information.
  • An example of the screen is shown in FIG. 5A.
  • the information processing device 100 can display graph data on the screen. This makes it easier for the user to consider whether to update single staining spectrum information.
  • the graph data is described above with reference to FIG. 4C, the type of graph data is not limited thereto.
  • the information processing apparatus 100 when updating the single staining spectrum information of PerCP-Cy5.5, the user selects the "Update" button 301. If not to update, the "Do not update” button 302 is selected. In response to the button 301 being selected, the information processing apparatus 100 changes the single staining spectrum information of PerCP-Cy5.5 to the latest information. In response to the button 302 being selected, the information processing apparatus 100 leaves the PerCP-Cy5.5 single staining spectrum information as it was. Furthermore, if there are other fluorescent dyes that the user should select to update, a screen related to the fluorescent dyes can be displayed. Alternatively, on one screen, the user may be asked whether or not to update all of the fluorescent dyes that the user should select whether to update. When similar processing is completed for all of the fluorescent dyes displayed as alert targets, the information processing device 100 advances the processing to step S17.
  • the information processing device 100 displays, for example, the original single staining spectrum information (Original, which is the first single staining spectrum information) and the latest single staining spectrum information (Latest, which is the second single staining spectrum information) on the screen 310.
  • single staining spectrum information can be displayed.
  • these two spectra are shown in one graph 311 in a superimposed manner.
  • the vertical axis is the fluorescence intensity expressed in arbitrary units
  • the horizontal axis is the channel number of the detector that detects the fluorescence. In this way, the number of each detector may be taken as an axis.
  • the screen 320 shown in the figure may include, for example, a graph 322 representing the above-mentioned change amount data in addition to a graph 321 in which the original single staining spectrum information and the latest single staining spectrum information are displayed in a superimposed manner.
  • the graph representing the change amount data may be, for example, a graph in which the vertical axis is the change amount and the horizontal axis is the channel number of the fluorescence detector, but is not limited thereto. Note that although the screen includes a graph representing the amount of change data, it may not include a graph representing a single staining spectrum.
  • step S17 the information processing device 100 ends the process of detecting a change in the single staining spectrum information.
  • the information processing apparatus 100 may perform, for example, panel design processing or unmixing processing. In these processes, the updated single staining spectrum is utilized in the process of detecting the change.
  • the biological sample analyzer may execute the BR>A unmixing process using the single staining spectrum information updated according to the present disclosure.
  • a biological sample analyzer 6100 shown in FIG. 6 includes a light irradiation unit 6101 that irradiates light to a biological sample S flowing through a flow path C, and a detection unit 6102 that detects light generated by irradiating the biological sample S with light. , and an information processing unit 6103 that processes information regarding the light detected by the detection unit.
  • Examples of the biological sample analysis device 6100 include a flow cytometer and an imaging cytometer.
  • the biological sample analyzer 6100 may include a sorting section 6104 that sorts out specific biological particles P in the biological sample.
  • An example of the biological sample analyzer 6100 including the sorting section is a cell sorter.
  • the biological sample S may be a liquid sample containing biological particles.
  • the biological particles are, for example, cells or non-cellular biological particles.
  • the cells may be living cells, and more specific examples include blood cells such as red blood cells and white blood cells, and reproductive cells such as sperm and fertilized eggs. Further, the cells may be directly collected from a specimen such as whole blood, or may be cultured cells obtained after culturing. Examples of the non-cellular biological particles include extracellular vesicles, particularly exosomes and microvesicles.
  • the biological particles may be labeled with one or more labeling substances (for example, a dye (particularly a fluorescent dye), a fluorescent dye-labeled antibody, etc.). Note that the biological sample analyzer of the present disclosure may analyze particles other than biological particles, and beads or the like may be analyzed for calibration or the like.
  • the flow path C is configured so that the biological sample S flows therethrough.
  • the flow path C may be configured such that a flow in which biological particles contained in the biological sample are arranged substantially in a line is formed.
  • the channel structure including the channel C may be designed so that laminar flow is formed.
  • the channel structure is designed such that a laminar flow is formed in which the flow of the biological sample (sample flow) is surrounded by the flow of the sheath liquid.
  • the design of the channel structure may be appropriately selected by those skilled in the art, and a known design may be adopted.
  • the flow channel C may be formed in a flow channel structure such as a microchip (a chip having a flow channel on the order of micrometers) or a flow cell.
  • the width of the channel C may be 1 mm or less, particularly 10 ⁇ m or more and 1 mm or less.
  • the channel C and the channel structure including the channel C may be formed from a material such as plastic or glass.
  • the biological sample analyzer of the present disclosure is configured so that the biological sample flowing in the flow path C, particularly the biological particles in the biological sample, is irradiated with light from the light irradiation unit 6101.
  • the biological sample analyzer of the present disclosure may be configured such that the interrogation point of light on the biological sample is in a channel structure in which the channel C is formed, or the interrogation point of the light The points may be configured to be outside the channel structure.
  • An example of the former is a configuration in which a channel C in a microchip or a flow cell is irradiated with the light.
  • the biological particles may be irradiated with the light after exiting the flow path structure (particularly its nozzle portion), and an example of this may be a jet-in-air type flow cytometer.
  • the light irradiation unit 6101 includes a light source unit that emits light and a light guide optical system that guides the light to an irradiation point.
  • the light source section includes one or more light sources.
  • the type of light source is, for example, a laser light source or an LED.
  • the wavelength of light emitted from each light source may be any wavelength of ultraviolet light, visible light, or infrared light.
  • the light guide optical system includes optical components such as a beam splitter group, a mirror group, or an optical fiber. Further, the light guide optical system may include a lens group for condensing light, and includes, for example, an objective lens. There may be one or more irradiation points where the biological sample and the light intersect.
  • the light irradiation unit 6101 may be configured to collect light irradiated from one or a plurality of different light sources onto one irradiation point.
  • the detection unit 6102 includes at least one photodetector that detects light generated by irradiating biological particles with light.
  • the light to be detected is, for example, fluorescence or scattered light (eg, any one or more of forward scattered light, back scattered light, and side scattered light).
  • Each photodetector includes one or more light receiving elements, such as a light receiving element array.
  • Each photodetector may include one or more photomultiplier tubes (PMTs) and/or photodiodes such as APDs and MPPCs as light receiving elements.
  • the photodetector includes, for example, a PMT array in which a plurality of PMTs are arranged in a one-dimensional direction.
  • the detection unit 6102 may include an imaging device such as a CCD or CMOS.
  • the detection unit 6102 can acquire images of biological particles (for example, bright field images, dark field images, fluorescence images, etc.) using the image sensor.
  • the detection unit 6102 includes a detection optical system that causes light of a predetermined detection wavelength to reach a corresponding photodetector.
  • the detection optical system includes a spectroscopic section such as a prism or a diffraction grating, or a wavelength separation section such as a dichroic mirror or an optical filter.
  • the detection optical system separates light generated by, for example, irradiating the biological particles with light, and allows the separated light to be detected by a plurality of photodetectors that exceed the number of fluorescent dyes labeled with the biological particles. configured.
  • a flow cytometer including such a detection optical system is called a spectral flow cytometer.
  • the detection optical system separates light corresponding to the fluorescence wavelength range of a specific fluorescent dye from the light generated by irradiating the biological particles with light, for example, and causes the corresponding photodetector to detect the separated light. It is configured like this.
  • the detection unit 6102 may include a signal processing unit that converts the electrical signal obtained by the photodetector into a digital signal.
  • the signal processing section may include an A/D converter as a device that performs the conversion.
  • a digital signal obtained by conversion by the signal processing section can be transmitted to the information processing section 6103.
  • the digital signal can be handled by the information processing unit 6103 as data related to light (hereinafter also referred to as "optical data").
  • the optical data may include, for example, fluorescence data. More specifically, the light data may be light intensity data, and the light intensity may be light intensity data of light including fluorescence (which may include feature quantities such as Area, Height, and Width). good.
  • the information processing unit 6103 includes, for example, a processing unit that processes various data (for example, optical data) and a storage unit that stores various data.
  • the processing unit acquires light data corresponding to a fluorescent dye from the detection unit 6102
  • the processing unit can perform fluorescence leakage correction (compensation processing) on the light intensity data.
  • the processing unit performs fluorescence separation processing on the optical data and obtains light intensity data corresponding to the fluorescent dye.
  • the fluorescence separation process may be performed, for example, according to the unmixing method described in JP-A No. 2011-232259.
  • the processing unit may acquire morphological information of the biological particles based on the image acquired by the imaging device.
  • the storage unit may be configured to store the acquired optical data.
  • the storage unit may further be configured to store spectral reference data used in the unmixing process.
  • the information processing section 6103 can determine whether to sort biological particles based on optical data and/or morphological information. Then, the information processing unit 6103 controls the sorting unit 6104 based on the result of the determination, so that the sorting unit 6104 can sort out biological particles.
  • the information processing unit 6103 may be configured to be able to output various data (for example, optical data and images). For example, the information processing unit 6103 can output various data (eg, two-dimensional plot, spectrum plot, etc.) generated based on the optical data. Further, the information processing unit 6103 may be configured to be able to accept input of various data, for example, accept gating processing on a plot by a user.
  • the information processing unit 6103 can include an output unit (for example, a display) or an input unit (for example, a keyboard) for executing the output or input.
  • the information processing unit 6103 may be configured as a general-purpose computer, and may be configured as an information processing device including, for example, a CPU, RAM, and ROM.
  • the information processing unit 6103 may be included in the casing in which the light irradiation unit 6101 and the detection unit 6102 are provided, or may be located outside the casing. Further, various processes or functions by the information processing unit 6103 may be realized by a server computer or cloud connected via a network.
  • the sorting unit 6104 performs sorting of biological particles according to the determination result by the information processing unit 6103.
  • the separation method may be a method in which droplets containing biological particles are generated by vibration, an electric charge is applied to the droplets to be separated, and the direction of movement of the droplets is controlled by electrodes.
  • the method of fractionation may be a method in which the traveling direction of the biological particles is controlled within the flow path structure and the fractionation is performed.
  • the flow path structure is provided with a control mechanism using, for example, pressure (injection or suction) or electric charge.
  • An example of the channel structure is a chip having a channel structure in which the channel C branches downstream into a recovery channel and a waste liquid channel, and specific biological particles are collected into the recovery channel. (For example, the chip described in JP-A No. 2020-76736).
  • the information processing device may execute the process of detecting the change described above using the single staining spectrum information acquired by the analysis process by the biological sample analyzer described above.
  • the biological sample analysis device described above may be configured as an information processing device according to the present disclosure.
  • the information processing unit 6103 may function as the processing unit 101 according to the present disclosure, and may be configured to execute the processing described above, for example.
  • the information processing device may execute the single staining spectrum information change detection process described above, and may execute panel design processing using the single staining spectrum information updated by the process. .
  • panel design process updated single staining spectral information is used, so better combinations of fluorescent reagents are proposed.
  • the panel design process may include a process of generating information regarding the correlation between single staining spectral information of two different fluorescent dyes (or fluorescent dye-labeled antibodies).
  • the information regarding the correlation is, for example, a correlation coefficient between single staining spectra or a square value of the correlation coefficient.
  • the correlation coefficient may be, for example, a Pearson correlation coefficient, a Spearman correlation coefficient, or a Kendall correlation coefficient.
  • the square value of the correlation coefficient can be used as an index of the degree of similarity between the two spectra, and the smaller the square value, the less similar the two single staining spectra are.
  • the information processing device can calculate the square value of the correlation coefficient using the updated single staining spectrum information as shown above, and use the calculated square value as an index to identify the combination of fluorescent dyes.
  • the identifying process may be performed such that the square value calculated for the two fluorescent dyes included in the combination becomes smaller.
  • the Pearson correlation coefficient can be calculated between two fluorescence spectra X and Y as follows.
  • fluorescence spectra X and Y can be expressed, for example, as follows.
  • Fluorescence spectrum X (X 1 , X 2 , ..., X 320 )
  • mean value ⁇ x
  • standard deviation ⁇ x (where X 1 to The average value ⁇ x is the average value of these fluorescence intensities.
  • the standard deviation ⁇ x is the standard deviation of these fluorescence intensities.
  • the standard deviation ⁇ x is the standard deviation of these fluorescence intensities.
  • the numerical value "320" is a value set for convenience of explanation, and the numerical value used in calculating the correlation coefficient is not limited to this value. The numerical value may be changed as appropriate depending on the configuration of the fluorescence detector, such as the number of PMTs (photomultiplier tubes) used for fluorescence detection.
  • Z Xn (n is 1 to 320) is the standardized fluorescence intensity and is expressed as follows.
  • Zx1 (X 1 - ⁇ x ) ⁇ x
  • Zx2 (X 2 - ⁇ x ) ⁇ x
  • ...Zx320 (X 320 - ⁇ x ) ⁇ x
  • Z Yn (n is 1 to 320) is also represented as follows.
  • Zy1 (Y 1 - ⁇ y ) ⁇ y
  • Zy2 (Y 2 - ⁇ y ) ⁇ y
  • ...Zy320 (Y 320 - ⁇ y ) ⁇ y
  • N is the number of data.
  • the information processing device executes the single staining spectrum information change detection process described above, and uses the single staining spectrum information updated by the process to perform fluorescence correction processing (particularly unmixing processing).
  • fluorescence correction processing particularly unmixing processing.
  • more accurate fluorescence correction is possible by using the updated single staining spectrum information.
  • the present disclosure also provides an information processing system including the processing unit described in "1. First Embodiment (Information Processing Apparatus)" above.
  • the information processing system may include the storage unit, input unit, output unit, and communication unit described in “1. First Embodiment (Information Processing Apparatus)” above. These components may be provided in one device or may be distributed and provided in multiple devices.
  • the information processing system may perform the processing related to single staining spectrum information described in "1. First embodiment (information processing device)”.
  • the information processing system 400 shown in the figure includes an information processing device 401, terminals 402-1 to 402-3, and a biological sample analysis device 403. These devices included in the information processing system 400 may be connected via a network 404.
  • the information processing device 401 may be configured as described in “1. First Embodiment (Information Processing Device)” above.
  • Information processing device 401 may be configured as a server computer, for example.
  • the information processing device 401 may be configured to collect single staining spectrum information acquired in analysis processing by the biological sample analysis device 403.
  • the biological sample analyzer 403 may be configured as described in "1. First Embodiment (Information Processing Apparatus)" above, and may be, for example, a flow cytometer. Biological sample analyzer 403 may be a closed cell sorter. The biological sample analyzer can perform an analysis process using the fluorescent dye. The number of biological sample analyzers that can be connected to the information processing system 400 is not limited to one shown in the figure, but may be one or more, for example. The biological sample analyzer 403 transmits the single staining spectrum of each fluorescent dye (or each fluorescent dye-labeled antibody) to the information processing device 401 or the terminals 402-1 to 402-3, for example, in response to the execution of analysis processing by the device. and the transmission may be performed automatically.
  • the terminals 402-1 to 402-3 are devices operated by users who execute information processing according to the present disclosure, and the terminals may be configured as information processing devices.
  • the information processing device may be configured as described in “1. First Embodiment (Information Processing Device)” above, and may be a desktop, laptop, or tablet terminal. The number of terminals is not limited to three as shown in the figure, and may be one or more, for example.
  • the information processing device 401 executes the single staining spectrum change detection process described in “1. First Embodiment (Information Processing Device)” above, and can display various screens generated along with the processing.
  • any of the terminals 402-1 to 402-3 executes the single staining spectrum change detection process described in "1. First embodiment (information processing device)" above, and further performs the Various screens generated along with the processing may be displayed.
  • the information processing method may include a process of detecting a change in single staining spectrum information of a fluorescent dye.
  • the processing is as described in 1. above. It may be performed as described in 1. above, in particular. (3). By performing this processing, appropriate single staining spectrum information can be selected, and thereby more accurate unmixing processing or panel design processing can be performed.
  • the present disclosure is based on the above 3. Also provided is a program for causing an information processing device to execute the information processing method described in .
  • the information processing method is as described in 1. above. and 3. The same description applies to this embodiment as well.
  • the program according to the present disclosure may be recorded, for example, on the above-mentioned recording medium, or may be stored in the above-mentioned information processing device or a storage unit included in the information processing device.
  • An information processing device comprising a processing section configured to detect a change in single staining spectrum information of a fluorescent dye.
  • the processing unit according to [1] wherein the processing unit compares first single staining spectrum information and second single staining spectrum information of the fluorescent dye, and determines whether the single staining spectrum information has changed based on the comparison.
  • Information processing device [3] The information processing device according to [2], wherein the first single staining spectrum information and the second single staining spectrum information are single staining spectrum information regarding the same fluorescent dye, but obtained at different times.
  • the information processing device according to any one of [1] to [3], wherein the single staining spectrum information is fluorescence intensity spectrum data of fluorescence generated by light irradiation to the fluorescent dye.
  • the single staining spectrum information is spectrum data represented by fluorescence intensity at each wavelength in a predetermined wavelength range.
  • the processing unit acquires change amount data representing a change between the first single staining spectrum information and the second single staining spectrum information of the fluorescent dye, and based on the change amount data and the comparison.
  • the information processing device according to [2], which determines whether single staining spectrum information has changed.
  • the information processing device according to any one of [8] to [10], wherein the information regarding the change includes single staining spectrum graph data of a fluorescent dye whose single staining spectrum information has changed.
  • the processing unit outputs a screen including a display prompting the user to select whether to update single staining spectrum information.
  • the processing unit is configured to execute panel design processing using updated single staining spectrum information.
  • the processing unit is configured to perform unmixing processing using updated single staining spectrum information.
  • An information processing system including a processing unit configured to detect a change in single staining spectral information of a fluorescent dye.
  • an information processing device having the processing section; A biological sample analyzer, The information processing system according to [15], comprising: [17] The information processing system according to [16], wherein the biological sample analyzer executes analysis processing using the fluorescent dye.
  • the biological sample analyzer is configured to transmit a single staining spectrum of the fluorescent dye obtained by the analysis process to the information processing device.
  • Information processing device 101 Processing section 102 Storage section 103 Input section 104 Output section 105 Communication section

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Abstract

L'objectif de la présente invention est de fournir une technologie permettant de déterminer de manière appropriée un état d'un réactif fluorescent. La présente divulgation concerne un dispositif de traitement d'informations pourvu d'une unité de traitement configurée pour détecter un changement dans des informations spectrales de coloration unique d'un colorant fluorescent. L'unité de traitement peut comparer des premières informations spectrales de coloration unique et des secondes informations spectrales de coloration unique du colorant fluorescent, et déterminer si les informations spectrales de coloration unique ont changé, sur la base de la comparaison. Les premières informations spectrales de coloration unique et les secondes informations spectrales de coloration unique sont des informations spectrales de coloration unique relatives au même colorant fluorescent, mais peuvent être acquises à différents moments. Les informations spectrales de coloration unique peuvent être des données spectrales d'intensité de fluorescence de fluorescence générée par émission de lumière sur le colorant fluorescent.
PCT/JP2023/007273 2022-03-10 2023-02-28 Dispositif de traitement d'informations et système de traitement d'informations WO2023171463A1 (fr)

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JP2002274813A (ja) * 2001-03-13 2002-09-25 Fuji Photo Film Co Ltd 一重項酸素発生方法及び一重項酸素検出方法
JP2011095073A (ja) * 2009-10-29 2011-05-12 Olympus Corp 撮像装置、及び、画像生成方法
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